Melanoma differentiation associated gene-5 and promoter and uses thereof

Abstract

The invention provides for an isolated nucleic acid encoding Mda-5 (melanoma differentiation associated gene-5) and an isolated Mda-5 polypeptide. The invention further provides a vector comprising the nucleic acid encoding Mda-5, as well as a host cell comprising the vector. The invention provides an antibody which specifically binds to an Mda-5 polypeptide. The invention further provides a method for determining whether a compound is an inducer of Mda-5 gene expression and assays to determine whether a compound modifies the enzymatic activity of the Mda-5 polypeptide.

1. An isolated nucleic acid comprising a nucleic acid having the sequence of the Melanoma Differentiation Associated Gene-5 (mda-5) promoter of SEQ ID NO:3, and having interferon beta-inducible promoter activity. 
     
     
       2. An isolated nucleic acid comprising a nucleic acid having a sequence which is at least 90% homologous to SEQ ID NO:3 and which has interferon beta-inducible promoter activity. 
     
     
       3. A vector comprising the isolated nucleic acid of any one of  claim 1  or  2 . 
     
     
       4. The vector of  claim 3 , wherein the isolated nucleic acid is operably linked to a therapeutic gene. 
     
     
       5. The vector of  claim 4 , wherein the therapeutic gene is a suicide gene. 
     
     
       6. The vector of  claim 5 , wherein the suicide gene is selected from
 a group consisting of a gene encoding an enzyme, an oncogene, a tumor suppressor gene, 
 a gene encoding a toxin, a gene encoding a cytokine, and a gene encoding oncostatin. 
 
     
     
       7. The vector of  claim 6 , wherein the gene encoding an enzyme is selected from a group consisting of the herpes simplex virus thymidine kinase gene, the  Escherichia coli  xanthine-guanine phosphoribosyltransferase gene, the  Escherichia coli  cytosine deaminase gene, and the  Escherichia coli  hypoxanthine phosphoribosyltransferase gene. 
     
     
       8. The vector of  claim 6 , wherein the oncogene is selected from a group consisting of the neu gene, the epidermal growth factor (EGF) gene, the ras gene, and the p53 gene. 
     
     
       9. The vector of  claim 6 , wherein the tumor suppressor gene is selected from a group consisting of the retinoblastoma (Rb) tumor suppressor gene, the Wilm's tumor (WT-1) gene, the phosphotyrosine phosphatase (PTPase) gene and the nm23 gene. 
     
     
       10. The vector of  claim 6 , wherein the gene encoding the toxin is selected from a group consisting of the gene encoding the  Pseudomonas aeruginosa  exotoxin A, the gene encoding the  Pseudomonas aeruginosa  exotoxin S, the gene encoding the  Corynebacterium diphtheriae  diphtheria toxin, the gene encoding the  Escherichia coli  LT toxin, the gene encoding the  Escherichia coli  Shiga toxin, the gene encoding the  Escherichia coli  Shiga-like toxin 1, the gene encoding the  Escherichia coli  Shiga-like toxin 2, the gene encoding the  Ricinus communis  toxin ricin, the gene encoding the  Abrus precatorius  toxin abrin and its variants, and the gene encoding the  Gelonium multiflorum  toxin gelonin. 
     
     
       11. The vector of  claim 6 , wherein the gene encoding the cytokine is selected from a group consisting of the gene encoding interferon α, the gene encoding interferon β, the gene encoding interferon γ, the gene encoding granulocyte/macrophage colony stimulating factor, the gene encoding tumor necrosis factor γ, and the gene encoding tumor necrosis factor α. 
     
     
       12. The vector of  claim 3 , wherein the vector is selected from a group consisting of an adenovirus, a retrovirus, an adeno-associated virus, a vaccinia virus, a herpes virus, an Epstein-Barr virus, a bacteriophage, a plasmid, a cosmid, or a replicon. 
     
     
       13. An isolated nucleic acid comprising a nucleic acid having the sequence of SEQ ID NO:25, and having interferon beta-inducible promoter activity. 
     
     
       14. An isolated nucleic acid comprising a nucleic acid having a sequence which is at least 90% homologous to SEQ ID NO:25 and which has interferon beta-inducible promoter activity. 
     
     
       15. A vector comprising the isolated nucleic acid of any one of  claim 13  or  14 . 
     
     
       16. The vector of  claim 15 , wherein the isolated nucleic acid is operably linked to a therapeutic gene. 
     
     
       17. The vector of  claim 15 , wherein the therapeutic gene is a suicide gene. 
     
     
       18. The vector of  claim 17 , wherein the suicide gene is selected from a group consisting of a gene encoding an enzyme, an oncogene, a tumor suppressor gene, a gene encoding a toxin, a gene encoding a cytokine, and a gene encoding oncostatin. 
     
     
       19. The vector of  claim 18 , wherein the gene encoding an enzyme is selected from a group consisting of the herpes simplex virus thymidine kinase gene, the  Escherichia coli  xanthine-guanine phosphoribosyltransferase gene, the  Escherichia coli  cytosine deaminase gene, and the  Escherichia coli  hypoxanthine phosphoribosyltransferase gene. 
     
     
       20. The vector of  claim 18 , wherein the oncogene is selected from a group consisting of the neu gene, the epidermal growth factor (EGF) gene, the ras gene, and the p53 gene. 
     
     
       21. The vector of  claim 18 , wherein the tumor suppressor gene is selected from a group consisting of the retinoblastoma (Rb) tumor suppressor gene, the Wilm's tumor (WT-1) gene, the phosphotyrosine phosphatase (PTPase) gene and the nm23 gene. 
     
     
       22. The vector of  claim 18 , wherein the gene encoding the toxin is selected from a group consisting of the gene encoding the  Pseudomonas aeruginosa  exotoxin A, the gene encoding the  Pseudomonas aeruginosa  exotoxin S, the gene encoding the  Corynebacterium diphtheriae  diphtheria toxin, the gene encoding the  Escherichia coli  LT toxin, the gene encoding the  Escherichia coli  Shiga toxin, the gene encoding the  Escherichia coli  Shiga-like toxin 1, the gene encoding the  Escherichia coli  Shiga-like toxin 2, the gene encoding the  Ricinus communis  toxin ricin, the gene encoding the  Abrus precatorius  toxin abrin and its variants, and the gene encoding the  Gelonium multaiflorum  toxin gelonin. 
     
     
       23. The vector of  claim 18 , wherein the gene encoding the cytokine is selected from a group consisting of the gene encoding interferon α, the gene encoding interferon β, the gene encoding interferon γ, the gene encoding granulocyte/macrophage colony stimulating factor, the gene encoding tumor necrosis factor γ, and the gene encoding tumor necrosis factor α. 
     
     
       24. The vector of  claim 15 , wherein the vector is selected from a group consisting of an adenovirus, a retrovirus, an adeno-associated virus, a vaccinia virus, a herpes virus, an Epstein-Barr virus, a bacteriophage, a plasmid, a cosmid, and a replicon. 
     
     
       25. The vector of  claim 4 , wherein the vector is selected from a group consisting of an adenovirus, a retrovirus, an adeno-associated virus, a vaccinia virus, a herpes virus, an Epstein-Barr virus, a bacteriophage, a plasmid, a cosmid, and a replicon. 
     
     
       26. The vector of  claim 16 , wherein the vector is selected from a group consisting of an adenovirus, a retrovirus, an adeno-associated virus, a vaccinia virus, a herpes virus, an Epstein-Barr virus, a bacteriophage, a plasmid, a cosmid, and a replicon.